Repositori institucional URV
| URV's Author/s: | ACERO SÁNCHEZ, JOSEP LLUÍS; HENRY ., OLIVIER; H. Joda; B. Werne Solnestam; L. Kvastad; E. Johansson; P. Akan; J. Lundeberg; N. Lladach; D. Ramakrishnan; I. Riley; O'SULLIVAN ., CIARA |
| Author, as appears in the article.: | J.L. Acero Sánchez; O.Y.F. Henry; H. Joda; B. Werne Solnestam; L. Kvastad; E. Johansson; P. Akan; J. Lundeberg; N. Lladach; D. Ramakrishnan; I. Riley; C.K. O'Sullivan |
| Author identifier: | n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; 0000-0002-0965-4655 |
| Journal publication year: | 2016 |
| Publication Type: | Article |
| ISSN: | 0956-5663 |
| Abstract: | Asymmetric multiplex ligation-dependent probe amplification (MLPA) was developed for the amplification of seven breast cancer related mRNA markers and the MLPA products were electrochemically detected via hybridization. Seven breast cancer genetic markers were amplified by means of the MLPA reaction, which allows for multiplex amplification of multiple targets with a single primer pair. Novel synthetic MLPA probes were designed to include a unique barcode sequence in each amplified gene. Capture probes complementary to each of the barcode sequences were immobilized on each electrode of a low-cost electrode microarray manufactured on standard printed circuit board (PCB) substrates. The functionalised electrodes were exposed to the single-stranded MLPA products and following hybridization, a horseradish peroxidase (HRP)-labelled DNA secondary probe complementary to the amplified strand completed the genocomplex, which was electrochemically detected following substrate addition. The electrode arrays fabricated using PCB technology exhibited an excellent electrochemical performance, equivalent to planar photolithographically-fabricated gold electrodes, but at a vastly reduced cost (>50 times lower per array). The optimised system was demonstrated to be highly specific with negligible cross-reactivity allowing the simultaneous detection of the seven mRNA markers, with limits of detections as low as 25 pM. This approach provides a novel strategy for the genetic profiling of tumour cells via integrated "amplification-to-detection". |
| Article's DOI: | 10.1016/j.bios.2016.04.018 |
| Link to the original source: | https://www.sciencedirect.com/science/article/pii/S0956566316302925?via%3Dihub |
| Paper version: | info:eu-repo/semantics/acceptedVersion |
| licence for use: | https://creativecommons.org/licenses/by/3.0/es/ |
| Department: | Enginyeria Química |
| Research group: | Group of Nanobiotechnology and Bioanalysis |
| Licence document URL: | https://repositori.urv.cat/ca/proteccio-de-dades/ |
| Thematic Areas: | Chemical engineering |
| Keywords: | Engineering controlled terms Electrochemical detection Engineering main heading |
| Entity: | Universitat Rovira i Virgili |
| Record's date: | 2016-05-03 |
| First page: | 224 |
| Last page: | 232 |
| Journal volume: | 82 |
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